Gastric H⁺/K⁺-ATPase Expression in Normal Laryngeal Tissue and Laryngeal Carcinoma

Figures & data

Table 1 Numbers of Specimens for Three Detection Methods

Total	Normal Laryngeal Tissues from 59 Patients			Laryngeal Carcinoma from 30 Patients			Paracarcinoma Tissues from 20 of 30 Patients
	195			90			60
	IHC	RT-PCR	WB	IHC	RT-PCR	WB	IHC	RT-PCR	WB
α	181	168	55	30	30	30	17	60	60
β	195	171	55	30	30	30	17	60	60

Table 2 The Expression of α-Subunit in Normal Laryngeal Tissues in Three Detection Methods

	IHC				RT-PCR			WB
	n	-(%)	+(%)	p	n	Relative Value (SD)	p	n	Relative Value (SD)	p
	181	136(75.1)	45(24.9)		168			55
Site
Glottic mucosa	12	11(91.7)	1(8.3)	0.18		0.955 ± 0.644	> 0.05		0.102 ± 0.019	> 0.05
Supraglottic mucosa	161	122(75.8)	39 (24.2)
Epiglottic	130	95(73.1)	35(26.9)	0.11		2.414 ± 0.349			0.200 ± 0.035
Ventricular fold	19	15(78.9)	4(21.1)			2.043 ± 0.476			0.233 ± 0.117
Arytenoid mucosa	12	12(100.0)	0(0.0)			2.000 ± 0.500			0.161 ± 0.029
Epiglottic cartilage	8	3(37.5)	5(62.5)			3.003 ± 1.020			0.307 ± 0.148

Abbreviations: IHC, immunohistochemical technique; SD, standard deviation; WB, Western blotting.

Figure 1 Expression levels of the α- and β-subunits of H⁺/K⁺-ATPases (proton pumps) in normal laryngeal tissues as revealed by immunohistochemical staining. (A) Gastric control tissues stained strongly for the α-subunit. (B) Gastric control tissues stained strongly for the α-subunit. (C) The α-subunit was expressed in normal epiglottic mucosa. (D) The α-subunit was expressed in normal epiglottic mucosa. (E) The α-subunit was expressed in normal false vocal cord mucosa. (F) The α-subunit was expressed in normal false vocal cord mucosa. (G) The α-subunit was expressed in normal epiglottic cartilage. (H) The α-subunit was expressed in normal epiglottic cartilage.

Table 3 The Expression of β-Subunit in Normal Laryngeal Tissues in Three Detection Methods

	IHC				RT-PCR			WB
	n	-(%)	+(%)	p	n	Relative Value (SD)	p	n	Relative Value (SD)	p
	195	152(77.9)	43(22.1)		171			55
Site
Glottic mucosa	11	11 (100.0)	0(0.0)	0.14		0.733 ± 0.306	> 0.05		0.096 ± 0.018	> 0.05
Supraglottic mucosa	175	139(79.4)	36 (20.6)
Epiglottic	137	107(78.1)	30(21.9)	0.13		2.352 ± 0.308			0.161 ± 0.029
Ventricular fold	24	18(75.0)	6(25.0)			2.909 ± 0.937			0.082 ± 0.004
Arytenoid mucosa	14	14(100.0)	0(0.0)			0.903 ± 0.349			0.082 ± 0.003
Epiglottic cartilage	9	2(22.2)	7(77.8)			1.903 ± 0.784			0.567 ± 0.133	< 0.05*

Notes: P< 0.05: A significant difference was detected between the expression of the β-subunit protein in the epiglottic cartilage and the other sites. *A significant difference was detected between the expression of the β-subunit protein in the epiglottic cartilage and the other sites.

Abbreviations: IHC, immunohistochemical technique; SD, standard deviation; WB, Western blotting.

Figure 2 Expression levels of mRNAs encoding the H⁺/K⁺-ATPase α- and β-subunits in normal laryngeal tissues and laryngeal carcinomas as revealed by real-time RT-PCR. (A) α-subunit mRNA, (B) β-subunit mRNA. *P < 0.05; **P < 0.01.

Figure 3 Expression levels of the H⁺/K⁺-ATPase α- and β-subunit proteins in representative normal laryngeal tissues and laryngeal carcinomas as revealed by Western blotting. (A) α-subunit protein, (B) β-subunit protein.

Notes: *P < 0.05; **P < 0.01.

Abbreviations: N, normal tissues; P, paracarcinoma tissues; T, laryngeal carcinoma tissues.

Table 4 The Relationship Between the Expression of α-Subunit or β-Subunits and Clinicopathological Features of Laryngeal Carcinoma

Clinicopatholical Parmeters	n	α-Subunit		p	β-Subunit		p
		-(%)	+(%)		-(%)	+(%)
Age
≥60	22	5(22.7)	17(77.3)	0.64	10(45.5)	12(54.5)	1.00
<60	8	3(37.5)	5(62.5)		3(37.5)	5(62.5)
Smoking
No	4	2(50.0)	2(50.0)	0.28	0(0.0)	4(100.0)	0.11
Yes	26	6(23.1)	20(76.9)		13(50.0)	13(50.0)
Alcohol
No	10	4(40.0)	6(60.0)	0.43	4(40.0)	6(60.0)	1.00
Yes	20	5(25.0)	15(75.0)		9(45.0)	11(55.0)
Tumor site
Supraglottis	5	2(40.0)	3(60.0)	0.89	3(60.0)	2(40.0)
Glottis	24	5(20.8)	19(79.2)		9(37.5)	15(62.5)
Subglottis	1	0(0.0)	1(100.0)		1(100.0)	0(0.0)
T
T1+T2	23	6(26.1)	17(73.9)	1.00	8(34.8)	15(65.2)	0.19
T3+T4	7	2(28.6)	5(71.4)		5(71.4)	2(28.6)
Lymph node metastasis
No	27	7(25.9)	20(74.1)		11(40.7)	16(59.3)	0.56
Yes	3	1(33.3)	2(66.7)		2(66.7)	1(33.3)
Clinical stage
I + II	23	6(26.1)	17(73.9)	1.00	8(34.8)	15(65.2)	0.19
III+ IV	7	2(28.6)	5(71.4)		5(71.4)	2(28.6)
Differentiation
Poorly	4	1(25.0)	3(75.0)	1.00	2(50.0)	2(50.0)	1.00
Moderately-well	26	7(26.9)	19(73.1)		11(42.3)	15(57.7)

Figure 4 Expression of the α-subunit of H⁺/K⁺-ATPases (proton pumps) in laryngeal carcinomas as revealed by immunohistochemical staining. (A) Gastric control tissues exhibited strong staining for the α-subunit. (B) The α-subunit was not expressed in paracarcinoma tissues. (C) The α-subunit was not expressed in laryngeal carcinomas. (D) Positive staining (brown) is evident in both the cytoplasm and plasma membrane of laryngeal carcinoma cells.

Figure 5 Expression of the β-subunit of H⁺/K⁺-ATPases (proton pumps) in laryngeal carcinomas as revealed by immunohistochemical staining. (A) Gastric control tissues exhibited strong staining for the β-subunit. (B). The β-subunit was not expressed in paracarcinoma tissues. (C) The β-subunit was not expressed in laryngeal carcinomas. (D) Positive staining (brown) is evident in both the cytoplasm and plasma membrane of laryngeal carcinoma cells.